Glutathione S-transferases of rabbit lung macrophages.
The catalytic activities of glutathione S-transferases (GSTs), particularly the alpha-class isozymes, can provide protection against oxidative stress through GSH-mediated metabolism of reactive products of lipid peroxidation. Lipid peroxidation products from oxidative metabolism in alveolar macrophages play an important role in mediating and regulating inflammatory response and injury in the lung. The rabbit has been used as an important animal model for studies of the role of alveolar macrophages in pulmonary pathology. Although rabbit lung macrophages display GST activity, the isozyme-specific expression of GSTs and the catalytic properties of these isozymes has not previously been defined. In present studies, we have purified the GST isozymes of rabbit alveolar macrophages obtained by bronchoalveolar lavage and performed immunologic and kinetic characterization of the purified isozymes. Results of our studies indicate the presence of three alpha-class isozymes (pI 10.2, 9.3, and 6.0) and one micro-class isozyme (pI 7.2). N-terminal sequence analysis of the micro-class isozyme indicated that it was distinct from the two previously described micro-class isozymes of rabbit. Kinetic studies indicated that two cationic alpha-class GSTs (pI 10.2 and 9.3) contribute the large majority of selenium independent GSH-peroxidase activity toward dilinoleoyl phosphatidylcholine hydroperoxide (kcat/Km values of 83.4 and 31.9 s-1 . M-1 . 10(3), respectively). A third alpha-class GST (pI 6.0) was shown to have highest catalytic activity toward conjugation of the 4-hydroxynonenal (4HNE) with GSH (kcat/Km = 1900 s-1 . M-1 . 10(3)). Structural and immunologic characterization of this GST isozyme indicated that it belongs to a subclass of the alpha-classGSTs selectively expressed in mesodermal origin cells that are exposed to high levels of oxidative stress and are characterized by high specific activity toward both lipid hydroperoxides and 4-HNE.
Morrison
R J
RJ
Department of Internal Medicine, The University of Texas Medical Branch, Galveston, Texas 77555, USA.
Singhal
S S
SS
Bidani
A
A
Heming
T A
TA
Awasthi
S
S
D000447:Aldehydes / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D018893:Bronchoalveolar Lavage; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D004137:Dinitrochlorobenzene / Q000378:metabolism; D005982:Glutathione Transferase / Q000737:chemistry / Q000302:isolation & purification*; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification*; D007700:Kinetics; D008054:Lipid Peroxides / Q000378:metabolism; D016676:Macrophages, Alveolar / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000378:metabolism; D011817:Rabbits
D000447:Aldehydes; D015853:Cysteine Proteinase Inhibitors; D004137:Dinitrochlorobenzene; D007527:Isoenzymes; D008054:Lipid Peroxides; D010713:Phosphatidylcholines; C002448:1,2-linoleoylphosphatidylcholine; D005982:Glutathione Transferase; C027576:4-hydroxy-2-nonenal